GSM6809981: JM8.N4 mESC, WT, RNAPolIIXChIP, rep2; Homo sapiens; Mus musculus; ChIP-Seq
Cells were fixed with 1% formaldehyde for 10 mins. Following quenching, chromatin was sonicated. Chromatin crosslinked to RPB1 was immunoprecipitated and then reverse crosslinked. Naked DNA was further sonicated to the 400-800bp range. DNA from RPB1 ChIP or input chromatin was used to prepare libraries using the NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB), using NEBNext Multiplex Oligos for Illumina for indexing samples. Libraries were amplified with a number of PCR cycles according to the manufacturer's protocol on the thermocycler using KAPA HiFi HotStart ReadyMix (Roche) and purified twice using AMPure XP beads (Beckman Coulter). Purified libraries were quantified using qPCR. Following qPCR quantification, libraries were submitted for sequencing.